Objective:To explore the clinical application value of serum N-glycan profiles for evaluating the severity of liver tissue inflammation in patients with chronic hepatitis B (CHB).Methods:A total of 221 CHB patients who underwent liver biopsy at Mengchao Hepatobiliary Hospital of Fujian Medical University from January 2018 to December 2020 were retrospectively enrolled. The Scheuer scoring system was used to assess the histological inflammation grade of the liver tissue. Serum N-glycan levels were measured using DNA sequencer-assisted N-glycan fingerprinting (NGFP). Using the upper limit of the alanine aminotransferase (ALT) reference value (40 U/L) as a cutoff, logistic regression models were developed to construct diagnostic models under two scenarios: normal ALT or abnormal ALT. Models based on serum N-glycan levels and serum N-glycan levels combined with routine laboratory indicators, were used to non-invasively evaluation of various pathological grades of liver tissue inflammation in CHB patients. The DeLong test was used to compare the diagnostic efficacy of the models by analyzing the areas under the receiver operating characteristic curve (AUC). Glycosylation-related gene expression differences associated with varying degrees of liver inflammation were analyzed using the Gene Expression Omnibus (GEO) database.Results:In CHB patients with normal ALT level, the relative abundances of N-glycan structure peak 1 (NGA2F) and peak 2 (NGA2FB) increased with higher liver inflammation grades, while the relative abundance of peak 5 (NA2) decreased ( P<0.05). The AUCs of the HIS-G model (HIS-G A) and its enhanced version (HIS-G A Plus) for identifying significant inflammation and necrosis (≥G2, indicating the initiation of antiviral therapy) were 0.805 (95% CI 0.690-0.899) and 0.904 (95% CI 0.821-0.960), respectively. In CHB patients with ALT>40 U/L, the relative abundances of peaks 1 (NGA2F), 2 (NGA2FB), and 3 (NG1A2F) increased with higher liver inflammation grades, while the relative abundances of peaks 8 (NA3) and 11 (NA4) decreased ( P<0.05). The AUCs of the HIS-G model (HIS-G B) and its enhanced version (HIS-G B Plus) for identifying significant inflammation (≥G2) were 0.810 (95% CI 0.727-0.889) and 0.838 (95% CI 0.754-0.901), respectively. With increasing liver inflammation grades, the expression levels of four glycosyltransferase genes (CHST4, FUT8, SLC51B, and ST8SIA4) were significantly upregulated ( P<0.05). Conclusions:Serum N-glycan biomarker models can be used to assist in evaluating the severity of liver tissue inflammation in CHB patients with both normal and abnormal ALT levels.

Objective:To explore the clinical application value of serum N-glycan profiles for evaluating the severity of liver tissue inflammation in patients with chronic hepatitis B (CHB).Methods:A total of 221 CHB patients who underwent liver biopsy at Mengchao Hepatobiliary Hospital of Fujian Medical University from January 2018 to December 2020 were retrospectively enrolled. The Scheuer scoring system was used to assess the histological inflammation grade of the liver tissue. Serum N-glycan levels were measured using DNA sequencer-assisted N-glycan fingerprinting (NGFP). Using the upper limit of the alanine aminotransferase (ALT) reference value (40 U/L) as a cutoff, logistic regression models were developed to construct diagnostic models under two scenarios: normal ALT or abnormal ALT. Models based on serum N-glycan levels and serum N-glycan levels combined with routine laboratory indicators, were used to non-invasively evaluation of various pathological grades of liver tissue inflammation in CHB patients. The DeLong test was used to compare the diagnostic efficacy of the models by analyzing the areas under the receiver operating characteristic curve (AUC). Glycosylation-related gene expression differences associated with varying degrees of liver inflammation were analyzed using the Gene Expression Omnibus (GEO) database.Results:In CHB patients with normal ALT level, the relative abundances of N-glycan structure peak 1 (NGA2F) and peak 2 (NGA2FB) increased with higher liver inflammation grades, while the relative abundance of peak 5 (NA2) decreased ( P<0.05). The AUCs of the HIS-G model (HIS-G A) and its enhanced version (HIS-G A Plus) for identifying significant inflammation and necrosis (≥G2, indicating the initiation of antiviral therapy) were 0.805 (95% CI 0.690-0.899) and 0.904 (95% CI 0.821-0.960), respectively. In CHB patients with ALT>40 U/L, the relative abundances of peaks 1 (NGA2F), 2 (NGA2FB), and 3 (NG1A2F) increased with higher liver inflammation grades, while the relative abundances of peaks 8 (NA3) and 11 (NA4) decreased ( P<0.05). The AUCs of the HIS-G model (HIS-G B) and its enhanced version (HIS-G B Plus) for identifying significant inflammation (≥G2) were 0.810 (95% CI 0.727-0.889) and 0.838 (95% CI 0.754-0.901), respectively. With increasing liver inflammation grades, the expression levels of four glycosyltransferase genes (CHST4, FUT8, SLC51B, and ST8SIA4) were significantly upregulated ( P<0.05). Conclusions:Serum N-glycan biomarker models can be used to assist in evaluating the severity of liver tissue inflammation in CHB patients with both normal and abnormal ALT levels.

BACKGROUND: Little was known about the association among time in range (TIR), time above range (TAR), time below range (TBR), and cancer mortality among patients with type 2 diabetes. We aimed to investigate the association among TIR, TAR, TBR, and the risk of cancer mortality among patients with type 2 diabetes. METHODS: A total of 6225 patients with type 2 diabetes were prospectively recruited in Shanghai, China. TIR was measured with continuous glucose monitoring at baseline and was defined as the average percentage of time in the target glucose range during a 24 h period. Cox proportion hazard regression analysis was used to determine the association between TIR and the risk of cancer mortality. RESULTS: During a mean follow-up of 7.10 years, we confirmed 237 death events related to cancer. The multivariable-adjusted hazard ratio (HR) for cancer mortality was 1.32 (95% confidence interval [CI]: 1.01-1.75) in patients with TIR ≤70% compared with those with TIR >70%. When TIR was considered as a continuous variable, the multivariable-adjusted HR for cancer mortality associated with each 10% decrease in TIR was 1.07 (95% CI: 1.02-1.14). In the site-specific analysis, a significant association between TIR as a continuous variable and the risk of hepatocellular cancer was found (HR: 1.24; 95% CI: 1.09-1.41). However, no relationship between hemoglobin A1c and cancer mortality was observed (HR: 1.04; 95% CI: 0.97-1.10). CONCLUSIONS The present study found an inverse association of TIR with the risk of cancer mortality among patients with type 2 diabetes. New evidence of TIR was added into the clinical practice that TIR may be an optimal target of glycemic control among patients with type 2 diabetes.
Blood Glucose ; Blood Glucose Self-Monitoring ; China ; Diabetes Mellitus, Type 2/complications* ; Humans ; Neoplasms ; Prospective Studies

Objective:To investigate the clinical management value of chitinase 3-like 1 protein(CHI3L1) in hepatocellular carcinoma (HCC) by studying the expression of CHI3L1 in peripheral blood, liver cancer and paired adjacent non-tumor tissues.Methods:Retrospective study. From 2013 to 2017, 405 patients with HCC in Third Affiliated Hospital of Naval Medical University were enrolled into the study. Meanwhile, 112 patients with liver cirrhosis (LC), 114 health subjects were included as disease and health controls. CHI3L1 in peripheral blood was detected by ELISA kit. Tissues array was made by collecting 90 pairs of tumor tissues and matched paracancer tissues, from HCC patients who were conformed by pathology. The expression of CHI3L1 in HCC tissues was analyzed by immunohistochemistry. Differences between independent groups were tested by Mann-Whitney U test or Kruskal Wallis H test, Pearson correlation analysis was used for analyzing the relationship between two subjects, and matched rank sum test was used for cancer tissue and adjacent tissue comparison. Results:The median (quartile) of CHI3L1 protein in LC group, HCC group and NC group was 195.8 (103.3,330.4) μg/L,118.2 (74.9,201.0) μg/L,46.8 (30.7,66.4) μg/L independently. The protein level of CHI3L1 in LC group was significantly higher than that in HCC group and health control group ( Z=5.186,12.928, P<0.001). HCC group was significantly higher than that in health control group ( Z=10.788, P<0.001). The level of CHI3L1 in HCC group was not related to whether liver cirrhosis was accompanied ( Z=-0.286, P=0.775). The level of serum CHI3L1 was positively correlated with noninvasive fibrosis markers (HA, PⅢNP, Ⅳ-C, FIB-4 index) ( r=0.202,0.159,0.299 and 0.221, P<0.05) and negatively correlated with ALB( r=-0.326, P<0.05) while positively correlated with AST and PT( r=0.138, 0.160, P<0.05). Positively correlation was observed between CHI3L1 and tumor size ( r=0.284, P<0.001). CNLC stage [CHI3L1 level in advanced group125.2(81.9,228.5)μg/L was higher than that in early group112.0(70.2,169.2)μg/L ( Z=-2.326, P=0.018)], but no correlation with microvascular invasion( Z=-1.531) and tumor capsule(χ 2=0.818, P>0.05). In 73 cases of HCC tissues, the positive rate of CHI3L1 was 78% (57/73) in cancer tissues and 83%(61/73) in paired adjacent non-tumor tissues. The staining intensity score of paracancer tissue 1.5(1.5,2.5) was higher than that of cancer tissue 1.5(1.5,2.0)( Z=-2.053, P=0.040). Conclusions:The tissue source of CHI3L1 protein in HCC includes cancer tissue and paracancerous tissue. The detection of serum CHI3L1 level is helpful to evaluate tumor load assessment and disease stratification management in HCC.

ORF3 protein, the single accessory protein encoded by porcine epidemic diarrhea virus (PEDV), is related to viral pathogenicity. In order to determine the cytoplasmic location signal of PEDV ORF3, we constructed a series of recombinant plasmids carrying full-length or truncated segments of PEDV DR13 ORF3 protein. When the acquired plasmids were transfected into Vero cells, expression and distribution of the EGFP-fused full-length ORF3 protein and its truncated forms in the cells were observed by laser confocal microscopy. The results showed that ORF3 protein or their truncated forms containing 40-91 aa segment including two transmembrane domains were localized in the cytoplasm, whereas ORF3 truncated peptides without the 40-91 aa segment were distributed in the whole cell (in both cytoplasm and nucleus). This suggests that the 40-91 aa is the key structural domain determining cytoplasmic location of PEDV ORF3 protein. The discovery provides reference for further clarifying intracellular transport and biological function of PEDV ORF3 protein.
Amino Acid Sequence ; Animals ; Chlorocebus aethiops ; Coronavirus Infections ; virology ; Cytoplasm ; virology ; Porcine epidemic diarrhea virus ; genetics ; Protein Domains ; Swine ; Vero Cells ; Viral Proteins ; chemistry ; metabolism

Objective To investigate the regulation of interferon α-1b (IFNα-1b) on protein kinase Cε(PKCε) and protein kinase Cα(PKCα) which inhibit the fibrosis of hepatic stellate cells (HSC) ,and to explore its mechanism .Methods HSC-T6 cells were treated with different levels of IFNα-1b (100 , 200 ,400 ,800 and 1000 U/mL) and the proliferation of HSC-T6 cells was analyzed by methyl thiazol tetrazolium (MTT) assay .Changes of hydroxyproline level were analyzed .The expressions of PKCεand PKCαwere detected by immunofluorescence staining . PKCε, PKCα,β-catenin and Survivin mRNA levels were detected by RT-PCR . PKCε, PKCα,β-catenin and Survivin protein levels were detected by Western blot . Variance analysis was conducted by using one-way ANOVA approach . Results The inhibition rates of 100 , 200 , 400 , 800 and 1000 U/mL IFNα-1b treatment after 24 hours of administration were (15 .85 ± 1 .05)％ ,(36 .59 ± 1 .03)％ ,(45 .12 ± 1 .05)％ ,(50 .00 ± 1 .01)％ and (62 .20 ± 1 .02)％ ,respectively ,with statistically significant differences among groups (F=27 .478 , P<0 .01) .The 48h inhibition rates were (20 .87 ± 1 .09)％ ,(43 .96 ± 1 .08)％ ,(53 .85 ± 1 .08)％ ,(64 .84 ± 1 .06)％ and (74 .72 ± 1 .07)％ ,respectively ,with statistically significant differences among groups (F=25 .321 , P< 0 .01 ) . half maximal inhibitory concentration at 48 h was 343 .47 U/mL . The levels of hydroxyproline in 100 ,200 and 400 U/mL IFNα-1b groups were (7 .48 ± 0 .28) ,(6 .26 ± 0 .17) and (3 .86 ± 0 .20) μg/mL ,respectively ,which were lower than that in control group (8 .47 ± 0 .32) μg/mL .The differences were all statistically significant (t=4 .033 ,10 .564 and 21 .160 ,respective ,all P<0 .05) .The fluorescence intensities of PKCεin 100 ,200 and 400 U/mL IFNα-1b groups were all lower than that of control group .The differences were statistically significant (t=1 .984 ,2 .457 and 7 .771 ,respectively ,all P<0 .05) .The fluorescence intensities of PKCαwere also significantly lower than that of control group (t=9 .232 ,15 .921 and 22 .222 ,respectively ,all P< 0 .01) .With the increase of IFNα-1b level ,the levels of HSC-T6 PKCε,PKCα,β-catenin and survivin were significantly lower than those of control group (t=7 .020 ,24 .562 ,45 .701 and 14 .241 ,respectively ,all P<0 .01) .With the increase of IFNα-1b ,the levels of HSC-T6 PKCε,PKCα,β-catenin and survivin were significantly lower than those of control group (t=9 .564 ,4 .409 ,10 .036 and 6 .794 ,respectively ,all P<0 .01) .Conclusions IFNα-1b can down-regulate the expression of collagen in hepatic stellate cells in a dose-dependent manner ,reduce the expressions of PKCε,PKCα,β-catenin and Survivin ,and inhibit the proliferation of HSC-T6 hepatic stellate cells .

Objective To investigate the clinical features of initially treated pulmonary tuberculosis(TB)patients with rifampin dependent mycobacterium tuberculosis Rv0341 antibody positive.Methods Forty two initially treated pulmonary TB patients with rifampin dependent mycobacterium tuberculosis Rv0341 antibody positive were included as the study group and 42 initially treated pulmonary TB patients with rifampin dependent mycobacterium tuberculosis Rv0341 antibody as the control group The aspects of clinical symptoms,X-ray imaging manifestations,positive rate in sputum culture of Mycobacterium tuberculosis and rifampin dependent mycobacterium tuberculosis,treatment effects,etc.were compared between the two groups.Results The incidence rates of expectoration,hemoptysis and shortness of breath in the study group were significantly higher than those in the control group(P＜0.05);the X-ray image manifestations in the study group showed that the lesions were much more involved in two lungs and multiple lung fields(P＜0.05),while thlesions in the control group more confined to single lung and one lung field(P＜ 0.05),moreover the rate appearing pulmonary cavity in the study group was higher than that in the control group(P＜0.05);the sputum culture positive rate of Mycobacterium tuberculosis had no statistical difference between the two groups(P＞0.05),but the sputum culture positive rate of rifampin dependent mycobacterium tuberculosis in the study group was significantly higher than that in the control group(P＜0.05);as for the treatment effects,the cure rate in the control group was higher than that in the study group(P＜0.05),the ineffective ratio in the control group was less than that in the study group.Conclusion Initially treated pulmonary TB patients with rifampin dependent mycobacterium tuberculosis Rv0341 antibody positive have the characteristics of severe clinical symptoms,wide range lesions,easily appearing cavity,high culture positive rate of rifampin dependent mycobacterium tuberculosis and poor treatment effect.

Objective To investigate the relationship between renal vascular resistance index( RI) and serum Apelin level in type 2 diabetic mellitus ( T2DM )?Methods Seventeen cases newly diagnosed T2DM patients with RI increased but without microalbuminuraia in Minhang Hospital Affiliated to Fudan University from December 2011 to December 2014 were selected as observation group,17 newly diagnosed T2DM patients with RI normal during the same period were selected as control group?Fasting plasma glucose(FPG),glycosylated hemoglobin A1C(HbAlC),blood lipids and fasting insulin(FINS),hepatic functional and renal function were tested in all the subjects?Serum Apelin level was detected by enzyme?linked immunosorbent assay?Results Compared with control group,serum Apelin level was significantly higher in observation group((179?2±122?4)μg/L vs?(56?7±50?6) μg/L,t=3?814,P<0?05)?Partial correlation analysis showed that the RI was positive correlated with Apelin ( r= 0?364, P= 0?040 )?Conclusion The serum Apelin levels elevated in newly diagnosed T2DM patients with RI increased, and RI is positively correlated with Apelin, which indicate that Apelin play an important role in the pathophysiology of early renal damage in patients with TsDM.

Objective To investigate the carriage and homology of carbapenemase genes of multidrug-resistant Acinetobacterbaumannii (MDRAB)in Wujiang area.Methods A total of 44 non-duplicated MDRAB isolated from patients in 3 general hospitals in Wujiang area from January 2010 to December 2013 were collected. Minimum inhibitory concentrations (MICs)were detected,carbapenemase genes OXA-51,OXA-23,OXA-24,OXA-58, IMP,TEM,SHV,and GES were amplified with polymerase chain reaction(PCR),homology of strains was detec-ted with pulsed-field gel electrophoresis (PFGE).Results 44 MDRAB strains were mainly collected from sputum (93.18% ),mainly distributed in intensive care unit (ICU),department of respiratory diseases,and department of neurosurgery,accounting for 45.45% ,27.27% ,and 13.64% respectively;MDRAB were both sensitive to minocy-cline and polymyxin B,resistance rates to piperacillin,ampicillin/sulbactam,ceftazidime,gentamicin,amikacin, and ciprofloxacin were all 100.00% ,resistance rates to imipenem and meropenem were both 97.73% . 44 MDRAB strains were all detected OXA-51,OXA-23 and TEM genes,12 strains were positive for GES gene,1 strain was positive for OXA-58 and SHV respectively,OXA-24 and IMP genes were not found ;MDRAB were divided into 7 types of G-A,which included 19,3,9,3,1,4,and 5 strains respectively,type A was mainly from two large gen-eral hospitals in Wujiang area (Wujiang First People’s Hospital and Shengze Hospital),type B,D and E strains were not detected in Wujiang First People’s Hospital,type E strain was only 1 isolate and was from Yongding Hos-pital,the other types were sporadically distributed.Conclusion Multidrug resistance of clinically isolated Acineto-bacterbaumannii is serious in Wujiang area,OXA-23 and TEM genes are major causes of multidrug resistance in Acinetobacterbaumannii,the main types are A and C,which present clonal spread.

Objective To study the correlation between preoperative dye exclusion test and liver function in patients with primary hepatic carcinoma.Methods This was a cross sectional survey.A total of 192 cases of primary liver cancer patients were recruited from May 2014 to March 2015 at the Second Military Medical University Affiliated Eastern Hepatobiliary Surgery Hospital.Hereinto, 160 cases were male and 32 females, the male to female ratio was 5: 1.The age of the patients ranged from 26 to 72 years old, and the average age was 50.5 years old.ICG 15 minutes retention rate of ICG clearance test was determined by PDD method in 192 cases of primary liver cancer patients.ICGR15 value was stratified into three stages: ICGR15 ＜ 10％ , ICGR1510％-20％ , and ICGR15 ＞ 20％.The ICGR15 stage of patients with different ChildPugh grades was analyzed.The biological liver function indexes of patients were simultaneous detected including TBIL, TBA, TP, ALB, PA, ALT, AST, PT-INR, HA, LN, Ⅲ, Ⅳ, APRI, PLT etc.The correlations of ICGR15 and biological indexes of liver function were analyzed using Spearman nonparametric correlation analysis.Results (1) ICGR15 was positively correlated with Child-Pugh grade (r =0.477, P ＜ 0.01) in the 192 cases of HCC.The hierarchical analysis showed that there were significant differences between ICGR15 and different Child-Pugh grades (P ＜ O.05).(2) Child-Pugh classification and ICGR15 comparison further showed that, ICGR15 increased with Child-Pugh grade.While ICG plasma clearance rate (ICGK) and effective hepatic blood flow (EHBF) reduced (P ＜ 0.05).(3) The correlation analysis between ICGR15 and biological indexes of liver function showed that: ICGR15 was positively correlated with TBIL,TBA, ALT, AST, AFU, GGT, PT-INR, HA, LN, Ⅲ, Ⅳ and APRI index [(AST/ULN) × 100/ PLT (× 109/L)] (r =0.422, 0.389, 0.219, 0.301, 0.219, 0.244, 0.325, 0.652,0.403, 0.523, 0.519, 0.434, P ＜ 0.05);and was negatively correlated with TP, ALB, PA, SOD, WBC, PLT (r =-0.290,-0.532, 0.546, 0.531, 0.256, 0.327, P＜ 0.05).Conclusions ICGR15 as a indicator for liver reserved and dynamic function can comprehensively reflect the liver reserve function is associated with the existing Child-Pugh grades and liver function biochemical indexes.Therefore, ICGR15 could be served as a sensitive index reflecting the preoperative liver reserve function.